Key Laboratory of Medical Diagnostics of Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, PR China.
Colloids Surf B Biointerfaces. 2013 Jan 1;101:481-6. doi: 10.1016/j.colsurfb.2012.07.021. Epub 2012 Jul 25.
A novel electrochemical immunosensor for detecting toxoplasma gondii-specific IgM (Tg-IgM) was constructed based on goldmag (Au-Fe(3)O(4)) nanoparticles and graphene sheets (GS). Thionine (Thi), as a mediator, was first electropolymerized on a nafion-GS (Nf-GS) modified electrode. Subsequently, gold nanoparticles (AuNPs) were attached onto the poly-thionine film through π-stacking interactions, and then were used to immobilize toxoplasma gondii antigen (Tg-Ag) for immunosensor fabrication. A sandwich-type immunoassay for Tg-IgM was performed using Au-Fe(3)O(4) labeled anti-IgM-horseradish peroxidase (HRP) as trace label. Electrochemical detection was carried out in the presence of H(2)O(2) as HRP substrate. Using Au-Fe(3)O(4) provided a simple, non-chemical damaging method for regeneration, and enhanced the HRP reduction ability toward H(2)O(2). The AuNPs/Thi/Nf-GS nanocomposite also had good conductivity and biocompatibility, which effectively improved the immunosensor sensitivity. Under optimal conditions, the immunosensor can detect Tg-IgM in two linear ranges from 0.0375 to 1.2 AU mL(-1) and from 2.0 to 18 AU mL(-1) with a detection limit of 0.016 AU mL(-1) (S/N=3). The immunosensor exhibited good reproducibility, stability, and selectivity as well.
一种基于金磁(Au-Fe(3)O(4))纳米粒子和石墨烯片(GS)的检测刚地弓形虫特异性 IgM(Tg-IgM)的新型电化学免疫传感器构建完成。首先,在 Nafion-GS(Nf-GS)修饰电极上电化学聚合了硫堇(Thi)作为媒介。随后,通过π-堆积相互作用将金纳米粒子(AuNPs)附着在聚硫堇薄膜上,然后用于固定刚地弓形虫抗原(Tg-Ag)以制备免疫传感器。采用 Au-Fe(3)O(4)标记的抗 IgM-辣根过氧化物酶(HRP)作为示踪标记,进行 Tg-IgM 的三明治型免疫分析。在 H(2)O(2)作为 HRP 底物的存在下进行电化学检测。使用 Au-Fe(3)O(4)提供了一种简单、非化学破坏性的再生方法,并增强了 HRP 对 H(2)O(2)的还原能力。AuNPs/Thi/Nf-GS 纳米复合材料还具有良好的导电性和生物相容性,有效提高了免疫传感器的灵敏度。在最佳条件下,该免疫传感器可以在两个线性范围内检测 Tg-IgM,范围从 0.0375 到 1.2 AU mL(-1)和 2.0 到 18 AU mL(-1),检测限为 0.016 AU mL(-1)(S/N=3)。该免疫传感器还表现出良好的重现性、稳定性和选择性。